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Characterization of a novel cytochrome P 450 from Amycolatopsis sp. CGMCC 1149 for hydroxylation of lovastatin
Author(s) -
Zong Hong,
Zhuge Bin,
Lu Xinyao,
Huo Xiaoyu,
Fang Huiying,
Song Jian,
Sun Jin
Publication year - 2014
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1002/bab.1247
Subject(s) - lovastatin , hydroxylation , reductase , biochemistry , chemistry , cytochrome , ferredoxin , cytochrome p450 , nicotinamide adenine dinucleotide phosphate , stereochemistry , enzyme , cholesterol , oxidase test
Wuxistatin, a novel and potent statin, is converted from lovastatin by A mycolatopsis sp. CGMCC 1149. In the bioconversion, lovastatin is firstly hydroxylated to 3‐hydroxymethyl lovastatin (product I) by a hydroxylase. In the current study, a novel hydroxylase gene p450lov was isolated from A mycolatopsis sp. CGMCC 1149 by degenerate PCR and self‐formed adaptor PCR and expressed in E scherichia coli . The gene encodes a 403‐amino‐acid protein with a molecular weight of 44.8 kDa and was designated as a new member of cytochrome P 450 ( CYP ) 105 family, CYP 105 A 44. Meanwhile, a lovastatin catalytic in vitro system was established, and an optimal hydroxylation reaction system contained 30 µM lovastatin, 600 µM NADH , 120 µM ferredoxin, 0.04 U ferredoxin−nicotinamide adenine dinucleotide phosphate reductase, and 100 µM CYP 105 A 44 in a final volume of 200 µL Tris HC l buffer (50 mM, pH 7.4). These would be helpful for further studies on the hydroxylation of statins.